Tenascin-X (TNX), a glycoprotein of the extracellular matrix (ECM), is expressed in various tissues and plays an important role in ECM architecture. The TNXB gene encoding TNX is known as the gene responsible for classic-like Ehlers-Danlos syndrome (cIEDS). To date, the role of TNX in dermal, muscular and obstetric features has been reported, but its role in bone homeostasis remains to be clarified. In this study, we found significant bone loss and upregulation of osteoclast marker gene expression in TNX-deficient mice. Further, TNX deficiency in the bone marrow promoted multinucleation of osteoclasts and resulted in increased bone resorption activity. These results indicate that multinucleated osteoclasts are the cause of bone loss in a TNX-deficient environment. Our findings provide new insight into the mechanism of osteoclast differentiation mediated by TNX and the pathology of cIEDS. (C) 2019 Elsevier Inc. All rights reserved.